Mechanism of Si Ni Decoction in Stabilizing Atherosclerotic Plaques in the Lower Extremities by Regulating Macrophage Polarization via NCOR1 Mitophagy
- 1Department of Vascular Diseases, Shanghai TCM‐Integrated Hospital Shanghai University of Traditional Chinese Medicine Shanghai China
- 2School of Pharmacy Macau University of Science and Technology. Avenida Wai Long Taipa China
Abstract
Introduction: Si Ni decoction (SND) is known for its anti‐inflammatory, immune‐regulating, and microcirculation‐improving effects. However, the molecular mechanisms underlying its role in atherosclerosis (AS) remain unclear. It aimed to explore the mechanisms by which SND stabilizes lower extremity atherosclerotic plaques through the regulation of macrophage polarization via mitophagy.
Materials and Methods: AS models were established in RAW 264.7 macrophages stimulated by lipopolysaccharide (LPS) and assigned into five groups [blank (BG), model (MG), low‐dose (LDG), medium‐dose (MDG), and high‐dose (HDG)]. The expression of pro‐inflammatory and anti‐inflammatory cytokines, M1 and M2 macrophage markers, mitophagy‐related proteins, and cell viability (CV) were detected in each group.
Result: Relative to the MG, in the LDG, MDG, and HDG, with increasing drug dosage, the levels of pro‐inflammatory cytokines interleukin‐6 (IL‐6) and tumor necrosis factor‐α (TNF‐α) decreased gradually, while the degree of IL‐10 raised; CD16/32 and COX2 were visibly reduced, and Arg1, CD206 visibly elevated; LC3, Parkin, FUNDC1 were visibly reduced, and P62 and CV were obviously elevated (p < 0.05). Meanwhile, SND dose‐dependently reversed the downregulation of NCOR1 expression induced by LPS (all p < 0.05).
Conclusion: The results of the present in vitro cell experiments showed that SND upregulated NCOR expression and improved abnormal mitophagy, thereby regulating macrophage polarization, alleviating inflammatory damage, and restoring mitochondrial function, thus providing preliminary in vitro experimental evidence for intervening in inflammatory lesions associated with atherosclerosis.
Aim: The present study aimed to investigate the in vitro mechanism by which SND regulates macrophage polarization through the mitophagy pathway, thereby elucidating its intervention effect on atherosclerosis‐related inflammatory damage. The specific objectives are as follows: (1) To reveal the potential mechanisms of SNT in regulating inflammatory responses by detecting the expression levels of pro‐inflammatory cytokines (such as IL‐6, TNF‐α) and anti‐inflammatory cytokines (such as IL‐10) in cells. (2) To further elucidate the regulatory effects of SNT on macrophage polarization by comparing the expression levels of M1‐type (such as CD16/32, COX2) and M2‐type (such as Arg1, CD206) macrophage markers after intervention with different doses of SNT. (3) To uncover the regulatory effects of SNT on macrophage function through the mitochondrial autophagy pathway by detecting the expression of mitochondrial autophagy markers (such as LC3, Parkin, P62, and FUNDC1) via Western blotting. (4) To assess the potential of SNT in improving mitochondrial function by measuring the OCR and ECAR of cells using the Seahorse XF real‐time cell analyzer. (5) To further evaluate the role of SNT in maintaining cell viability by detecting its effects on cell proliferation and survival rate using the MTS assay.
Core Tips: Atherosclerosis models were established in RAW 264.7 macrophages stimulated by LPS, and interventions with different doses of SND were applied. The experimental groups included the blank group, model group, and different doses of SND groups. Real‐time PCR was used to detect the gene expression of pro‐inflammatory cytokines (IL‐6, TNF‐α) and anti‐inflammatory cytokines (IL‐10); Western blotting was used to detect the expression of M1 and M2 macrophage markers and mitochondrial autophagy‐related proteins; the Seahorse analyzer was used to detect mitochondrial function; and the MTS method was used to assess cell viability. The present study suggests that SND attenuates inflammatory damage, restores mitochondrial function, and modulates macrophage polarization via regulation of NCOR1 and aberrant mitophagy at the in vitro cellular level. This study provides new molecular mechanisms for the potential application of SND in atherosclerosis and offers experimental evidence for clinical treatment.
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BibTeX
@article{liu2026mechanism,
title = {{Mechanism of Si Ni Decoction in Stabilizing Atherosclerotic Plaques in the Lower Extremities by Regulating Macrophage Polarization via NCOR1 Mitophagy}},
author = {Zhe Liu and Shuhui Ge and Jianfei Yang and Ying Zhou and Li Jin and Yudong Fang},
journal = {Immunity, Inflammation and Disease},
year = {2026},
month = oct,
doi = {10.1002/iid3.70546},
url = {https://doi.org/10.1002/iid3.70546},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Mechanism of Si Ni Decoction in Stabilizing Atherosclerotic Plaques in the Lower Extremities by Regulating Macrophage Polarization via NCOR1 Mitophagy
AU - Zhe Liu
AU - Shuhui Ge
AU - Jianfei Yang
AU - Ying Zhou
AU - Li Jin
AU - Yudong Fang
PY - 2026
DA - 2026/10/01
JO - Immunity, Inflammation and Disease
DO - 10.1002/iid3.70546
UR - https://doi.org/10.1002/iid3.70546
AB - Introduction: Si Ni decoction (SND) is known for its anti‐inflammatory, immune‐regulating, and microcirculation‐improving effects. However, the molecular mechanisms underlying its role in atherosclerosis (AS) remain unclear. It aimed to explore the mechanisms by which SND stabilizes lower extremity atherosclerotic plaques through the regulation of macrophage polarization via mitophagy. Materials and Methods: AS models were established in RAW 264.7 macrophages stimulated by lipopolysaccharide (LPS) and assigned into five groups [blank (BG), model (MG), low‐dose (LDG), medium‐dose (MDG), and high‐dose (HDG)]. The expression of pro‐inflammatory and anti‐inflammatory cytokines, M1 and M2 macrophage markers, mitophagy‐related proteins, and cell viability (CV) were detected in each group. Result: Relative to the MG, in the LDG, MDG, and HDG, with increasing drug dosage, the levels of pro‐inflammatory cytokines interleukin‐6 (IL‐6) and tumor necrosis factor‐α (TNF‐α) decreased gradually, while the degree of IL‐10 raised; CD16/32 and COX2 were visibly reduced, and Arg1, CD206 visibly elevated; LC3, Parkin, FUNDC1 were visibly reduced, and P62 and CV were obviously elevated (p < 0.05). Meanwhile, SND dose‐dependently reversed the downregulation of NCOR1 expression induced by LPS (all p < 0.05). Conclusion: The results of the present in vitro cell experiments showed that SND upregulated NCOR expression and improved abnormal mitophagy, thereby regulating macrophage polarization, alleviating inflammatory damage, and restoring mitochondrial function, thus providing preliminary in vitro experimental evidence for intervening in inflammatory lesions associated with atherosclerosis. Aim: The present study aimed to investigate the in vitro mechanism by which SND regulates macrophage polarization through the mitophagy pathway, thereby elucidating its intervention effect on atherosclerosis‐related inflammatory damage. The specific objectives are as follows: (1) To reveal the potential mechanisms of SNT in regulating inflammatory responses by detecting the expression levels of pro‐inflammatory cytokines (such as IL‐6, TNF‐α) and anti‐inflammatory cytokines (such as IL‐10) in cells. (2) To further elucidate the regulatory effects of SNT on macrophage polarization by comparing the expression levels of M1‐type (such as CD16/32, COX2) and M2‐type (such as Arg1, CD206) macrophage markers after intervention with different doses of SNT. (3) To uncover the regulatory effects of SNT on macrophage function through the mitochondrial autophagy pathway by detecting the expression of mitochondrial autophagy markers (such as LC3, Parkin, P62, and FUNDC1) via Western blotting. (4) To assess the potential of SNT in improving mitochondrial function by measuring the OCR and ECAR of cells using the Seahorse XF real‐time cell analyzer. (5) To further evaluate the role of SNT in maintaining cell viability by detecting its effects on cell proliferation and survival rate using the MTS assay. Core Tips: Atherosclerosis models were established in RAW 264.7 macrophages stimulated by LPS, and interventions with different doses of SND were applied. The experimental groups included the blank group, model group, and different doses of SND groups. Real‐time PCR was used to detect the gene expression of pro‐inflammatory cytokines (IL‐6, TNF‐α) and anti‐inflammatory cytokines (IL‐10); Western blotting was used to detect the expression of M1 and M2 macrophage markers and mitochondrial autophagy‐related proteins; the Seahorse analyzer was used to detect mitochondrial function; and the MTS method was used to assess cell viability. The present study suggests that SND attenuates inflammatory damage, restores mitochondrial function, and modulates macrophage polarization via regulation of NCOR1 and aberrant mitophagy at the in vitro cellular level. This study provides new molecular mechanisms for the potential application of SND in atherosclerosis and offers experimental evidence for clinical treatment.
SN - 2050-4527
ER -